c-Fos [pT325] Antibody | catalog# XBP-4057
Background
c-Fos is a ~60 kDa immediate early gene product and a member of AP-1 family. c-Fos dimerizes with c-Jun to form the transcriptionally active complex AP-1, and functions in regulating cell proliferation and differentiation. c-Fos expression and activation are induced in many tumors (including breast cancer), and in response to a variety of stimuli (including growth factors, cytokines and stress). ERK1&2 mediate growth factor-induced c-Fos induction and transcripional activation by directly phosphorylating the carboxyl terminal domain of c-Fos. c-Fos is phosphorylated on several serine and threonine sites, including threonine 325 and 232. Phosphorylation of threonine 325 is believed to augment c-Fos transcriptional activity.
Description
![c-Fos [pT325] Antibody](./image/product/20/25/XBP-4057.gif)
Lysates prepared from A431 cells left untreated (1) or treated with EGF (lanes 2 & 3) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were then left untreated (lanes 1 & 2) or treated with Lambda phosphatase (lane 3), blocked with a 3% Milk-TBST buffer for one hour at room temperature, and incubated with c-Fos [pT325] antibody for two hours at room temperature in a 3% Milk-TBST buffer. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal™ method.
Specifications
Formulation
Rabbit polyclonal immunoglobulin in Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol, with 1.0 mg/mL BSA (IgG, protease free) as a carrier.Clonality/Clone
This is a polyclonal antibody.Specificity
Human and mouse c-Fos. Rat c-Fos (100% homologous) has not been tested, but is expected to react.Source
c-Fos antibody was produced against a chemically synthesized phosphopeptide derived from the region of human c-Fos that contains threonine 325. The sequence is conserved in human, mouse and rat.Purification
c-Fos antibody was purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated c-Fos. The final product is generated by affinity chromatography using a c-Fos-derived peptide that is phosphorylated at threonine 325.Host
c-Fos [pT325] antibody was raised in rabbit.Please use anti-rabbit secondary antibodies.
Application
For Western blotting applications, we recommend using the antibody at a 1:1000 dilution.Tested Application
WBBuffer
Rabbit polyclonal immunoglobulin in Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol, with 1.0 mg/mL BSA (IgG, protease free) as a carrier.Storage
Store at -20˚C. We recommend a brief centrifugation before opening to settle vial contents. Then, apportion into working aliquots and store at -20˚C. For shipment or short-term storage (up to one week), 2 - 8˚C is sufficient.Positive Control
- Human epidermoid carcinoma (A431) cells stimulated with EGF or 15% serum.]
Species Reactivity
HReference
- Monje, P., et al. (2003) Phosphorylation of the carboxyl-terminal transactivation domain of c-Fos by extracellular signal-regulated kinase mediates the transcriptional activation of AP-1 and cellular transformation induced by platelet-derived growth factor. Mol. Cell. Biol. 23(19):7030-7043.
- Wang, L.H., et al. (2003) The cis decoy against the estrogen response element suppresses breast cancer cells via target disrupting c-fos not mitogen-activated protein kinase activity. Cancer Res. 63(9):2046-2051.
- Deng, T. and M. Karin (1994) c-Fos transcriptional activity stimulated by H-Ras-activated protein kinase distinct from JNK and ERK. Nature 371(6493):171-175.
- Lu, Y., et al. (2004) Overexpression of ErbB2 receptor inhibits IGF-I-induced Shc-MAPK signaling pathway in breast cancer cells. Biochem. Biophys. Res. Commun. 313(3):709-715.]